右归丸对糖尿病视网膜病变大鼠PI3K/Akt信号通路的影响

目的：探讨“阴中求阳”立法之右归丸对糖尿病视网膜病变大鼠PI3 K/Akt信号通路的影响。方法：SD大鼠80只,随机分组,10只为正常组,余大鼠通过一次性腹腔注射链脲佐菌素溶液(60 mg/kg )联合大鼠玻璃体腔注射VEGF(0.05μg)的方式建立糖尿病视网膜病变增殖期大鼠模型。模型建立成功后,将最终成模大鼠随机分为4组：右归丸高中低剂量组、模型组,右归丸高中低剂量组每天给予相应右归丸浓缩液浓度进行灌胃,模型组、正常组每天给予等剂量蒸馏水灌胃。连续灌胃3 mo后,采取SP免疫组织化学、Western-blot法分别观察检测大鼠视网膜组织PI3 K和Akt的表达情况。结果：免疫组织化学观察显示PI3 K和Akt免疫组化染色的阳性表达,在视网膜上均为棕黄色颗粒;正常组PI3 K和Akt表达部分分布于神经节细胞层,内核也有少量表达,呈弱阳性免疫反应;模型组、右归丸各治疗组大鼠视网膜神经节细胞层、内丛状层及内外颗粒层都有阳性表达;与模型组比较,右归丸中、高剂量组PI3 K和Akt表达均减弱,右归丸低剂量组表达减弱不明显;与右归丸低剂量组比较,右归丸中、高剂量组PI3 K和Akt表达减弱;右归丸中、高剂量组表达强弱比较无差异。 Western-blot检测结果显示与正常组比较,模型组、治疗组表达差异有显著统计学意义( P<0.01),模型组、治疗组PI3 K和Akt蛋白表达水平显著升高;与模型组比较,右归丸中、高剂量组表达差异有统计学意义(P<0.05),右归丸中、高剂量组PI3 K和Akt蛋白表达水平均降低,右归丸低剂量组表达差异无统计学意义(P>0.05);与右归丸低剂量组比较,右归丸中、高剂量组表达差异有统计学意义(P<0.05),右归丸中、高剂量组PI3 K和Akt表达水平下降;右归丸中、高剂量组表达比较,差异无统计学意义(P>0.05)。结论：基于“阴中求阳”立法之右归丸可以通过影响PI3 K和Akt蛋白表达水平,抑制PI3 K/Akt 信号通路活化,延缓糖尿病视网膜病变的病变进程,为糖尿病视网膜病变防盲治疗提出新的治疗思路与科学依据。     

大鼠实验性慢性高眼压对视网膜神经节细胞的损伤

目的:制作高眼压大鼠模型,观察高眼压对视神经的损害。方法:成年Wistar雄性大鼠65只,烧灼右眼上方2支和外侧1支巩膜上静脉,建立慢性高眼压模型。左眼作为对照眼。对造模成功的,分别于造模后1d,1,2,3,4,6,8,10wk各摘除6只大鼠双眼。在取出眼球前24h,用Fluoro-gold进行视网膜神经节细胞(retinal ganglion cells,RGC)逆行性染色,做视网膜铺片计数RGC,观察不同时段高眼压对RGC的影响。结果:右眼巩膜上静脉烧灼后各时间点造模眼平均眼压分别为42.2±1.8mmHg,37.9±2.3mmHg,36.1±2.0mmHg,33.6±2.2mmHg,32.2±2.4mmHg,30.1±2.0mmHg,30.5±2.1mmHg和27.6±1.3mmHg。术后各时间点的成模率分别为80.0%,76.9%,74.5%,71.7%,63.8%,56.1%,42.9%,41.4%。成模率和成模眼的眼压随时间延长呈下降趋势。实验组和对照组的RGC密度在早期(巩膜上静脉烧灼术后3wk内)没有显著差别。造模后4wk高眼压组RGC密度明显低于对照组(P<0.05),随着时间的推移差别越来越显著。结论:巩膜上静脉烧灼法能诱导出持续的肯定的大鼠慢性高眼压模型,成模眼的眼压和随时间而下降。高眼压持续的时间越长,RGC的损失越多。     

Colivelin对视神经保护作用的初步研究

目的初步探讨Colivelin对大鼠外伤性视神经损伤后的神经保护作用和机理,及其保护效果是否具有剂量依赖性。方法 40只健康2月龄Wistar大鼠,随机分为模型组、安慰剂组及Colivelin低剂量组(10-6 μmol·L-1 Colivelin)、Colivelin中剂量组(10-4μmol·L-1Colivelin)、Colivelin高剂量组(10-2 μmol·L-1 Colivelin),每组8只。40只大鼠均做单侧视神经损伤模型,模型组另一侧未做任何处理的8眼为空白对照组。应用无创血管夹建立大鼠视神经夹伤模型,造模后30 min Colivelin治疗组玻璃体内分别注射不同浓度Colivelin 5 μL,安慰剂组注射5μLPBS缓冲液,模型组和空白对照组不给予任何治疗。注药后7d,通过视网膜切片技术,进行HE染色观察视网膜神经节细胞(retinal ganglion cell,RGC)的形态及数目,TUNEL染色法检测RGC的凋亡以及免疫组织化学法检测视网膜中caspase-3的表达。结果 HE染色可见空白对照组视网膜各层细胞排列整齐密集,模型组、安慰剂组、Colivelin治疗组均可见部分RGC核固缩,但随着Colivelin注射浓度的增加,RGC发生核固缩的数量减少。RGC计数:空白对照组每400倍光镜视野下RGC数量为25.750±1.264,模型组为8.236±1.239,安慰剂组为8.514±1.222,Colivelin低剂量组为14.500±1.021,Colivelin中剂量组为16.250±1.319,Colivelin高剂量组为18.097±1.323,除模型组与安慰剂组之间差异无统计学意义(P>0.05)外,其他各组之间差异均有统计学意义(均为P<0.05)。TUNEL染色:模型组和安慰剂组注药后7 d TUNEL染色可见大量凋亡细胞,Colivelin治疗组凋亡细胞数量随Colivelin注射浓度的增加依次递减。RGC细胞凋亡率:模型组为(60.928±2.961)%,安慰剂组为(61.446±2.755)%,Colivelin低剂量组为(58.432±2.835)%,Colivelin中剂量组为(51.948±2.802)%,Colivelin高剂量组为(47.656±2.331)%。Caspase-3表达:模型组、安慰剂组及Colivelin低剂量组、中剂量组、高剂量组均可见caspase-3表达。平均光密度值显示,模型组和安慰剂组相比caspase-3表达无明显差异(分别为0.482±0.012和0.486±0.012),均高于Colivelin治疗组(均为P<0.05),并且Colivelin低剂量组、中剂量组、高剂量组组间差异亦均有统计学意义(分别为0.411±0.017、0.326±0.018、0.234±0.016;均为P<0.05)。结论 Colivelin能有效增加视神经损伤后RGC的数量,抑制其凋亡,减少caspase-3表达,且这一作用呈剂量依赖性。     

灯盏细辛对大鼠慢性高眼压模型视神经损伤保护的实验研究

目的:建立大鼠慢性高眼压模型,观察灯盏细辛(Erigeronbrevicapas hand mass,EBHM)对眼压升高诱导视神经损伤的保护作用。方法:选用健康成年Wistar大鼠90只,分为3组。用波长为532nm的氪离子黄绿激光光凝第1和第2组大鼠双眼小梁网,建立大鼠高眼压模型。在眼压升高后1wk开始用EBHM对第2组大鼠15mg/100g肌肉注射,行视神经保护性治疗。第1组作为光凝对照组,第3组大鼠作为正常对照组。在第9wk同时处死3组大鼠做全视网膜铺片,1%甲苯氨蓝染色,记数平均视网膜神经节细胞(retinalganglion cells,RGCs)密度。结果:所有光凝眼眼压均中等程度升高,光凝前眼压为14.70±3.2mmHg;光凝后第3,6,9wk眼压分别为27.25±4.75,28.75±6.24,25.47±5.60mmHg,与光凝前比较,差别有显著性(P<0.05)。经视网膜铺片甲苯胺蓝染色,视网膜RGCs密度值(个/mm2)为:第1组1654±136,第2组2135±125,第3组2516±196。第2组大鼠视网膜RGCs密度值与第1和第3组大鼠RGCs密度值比较,差别有显著性(P<0.05)。结论:光凝大鼠小梁网成功建立大鼠慢性高眼压模型,光凝眼眼压中等程度升高,RGC密度降低;EBHM能够部分保护大鼠慢性高眼压诱导的视神经损害。     

葛根素对大鼠视网膜神经节细胞的作用

目的探讨葛根素对大鼠视神经夹伤模型视网膜神经节细胞(retinal ganglion cells,RGC)的保护作用.设计随机对照实验研究.对象40只SD大鼠.方法将40只大鼠随机分5组,每组8只,分别为生理盐水组,莫尼定组,葛根素高、中、低剂量组.右眼行视神经夹伤,左眼为正常对照.术后24天双侧上丘荧光金逆行标记.术后28天视网膜定向铺片、拍摄荧光照片及图像分析.RGC的存活率为右眼RGC密度除以左眼RGC密度,再乘以100%.主要指标RGC的存活率.结果40只SD大鼠RGC存活率生理盐水组为(53.99±6.69)%,葛根素低剂量组(58.67±6.55)%,中剂量组(63.85±5.75)%,高剂量组(69.66±4.79)%,莫尼定组(62.10±3.90)%.葛根素高、中剂量组及莫尼定组均与生理盐水组比较有显著性差异(P值均小于0.05).结论葛根素中剂量组、高剂量组及莫尼定组对视神经损伤后RGC的存活有明显的增强作用.     

高良姜素对青光眼大鼠视网膜神经节细胞的保护作用

目的 研究高良姜素对青光眼大鼠视网膜神经节细胞(RGC)的保护作用。方法 取48只SD大鼠随机分为对照组、对照+高良姜素组、高眼压组和高眼压+高良姜素组,每组12只(12眼)。对照+高良姜素组和高眼压+高良姜素组大鼠接受高良姜素滴眼液治疗,对照组和高眼压组大鼠则接受相同剂量二甲基亚砜溶剂滴眼。于造模后第0～28天监测大鼠眼压,并于造模后第28天处死大鼠,收集各组大鼠右眼球,并采用HE染色观察大鼠视网膜形态变化,采用免疫组织化学染色分析各组大鼠视网膜中GFAP、Toll样受体4(TLR4)和NOD样受体3(NLRP3)表达。从4只4 d龄大鼠视网膜中分离RGC,分为空白组、光气组和CoCl₂+高良姜素组。光气组和光气+高良姜素组RGC与200μmol·L^-1 CoCl₂一起孵育48 h,光气+高良姜素组RGC中加入20μmol·L^-1高良姜素干预48 h。收集各组细胞,采用流式细胞术和Western blot检测RGC的凋亡率及相关蛋白的相对表达情况,并采用ELISA检测空白组、光气组和光气+高良姜素...     

大鼠慢性高眼压青光眼模型的构建及鉴定

目的:手术建立慢性高眼压青光眼大鼠模型对研究青光眼发病机制、治疗和视神经保护这三方面的研究尤为必要。方法:烧灼3条表浅巩膜静脉,并术中联合应用结膜瓣下浸有0.2g/L的丝裂霉素的凝胶海绵,阻断大鼠的眼部房水外流,建立大鼠慢性高眼压模型,手术后2h;1,3,7,14,28,56d处死5只大鼠,空白对照组亦处死5只大鼠;摘除眼球光镜下进行视网膜形态的观察及视网膜厚度测量,研究眼压、视网膜神经节细胞(retinal ganglion cells,RGC)和视神经损伤的特性。结果:大鼠手术眼与假手术眼、空白对照眼的术后各时间点的眼压比较差异有显著性(P<0.01)。结论:建立大鼠慢性高眼压模型操作简单,重复性好,成模率高,高眼压可稳定维持长时间,慢性高眼压将导致整个视网膜的变薄和萎缩。     

热休克蛋白27在实验性青光眼中表达的研究

目的　观察大鼠眼压升高后热休克蛋白 2 7(HSP2 7)在视网膜中的表达。方法　5 0只Wistar大鼠随机分为高眼压组和sham对照 (假手术 )组。采用电凝鼠巩膜表面至少3组静脉及角膜缘周围血管 ,减少房水静脉回流升高眼压。观察手术后 1、2、3、4及 8周大鼠眼压 ,同时免疫组化检测视网膜中HSP2 7的表达及分布情况。结果　高眼压组右眼术后眼压明显升高。术后 1周眼压 :(30 .12± 5 .18)mmHg(1kPa =7.5mmHg) ,1周后眼压基本稳定。术后各时间点高眼压组右眼眼压与术前、左眼及sham对照组右、左眼间比较 ,差异有显著性 (P <0 .0 0 1)。视网膜中HSP2 7阳性表达主要表现在视网膜神经节细胞的胞浆内及神经纤维层中 ,HSP2 7阳性表达率在术后 1、2、3、4及 8周 ,高眼压组右眼与左眼、sham对照组右、左眼间比较 ,差异有显著性 (P <0 .0 0 1) ,同时发现视网膜中HSP2 7阳性表达随着眼压升高及高眼压持续时间延长逐渐增强。结论　内源性HSP2 7在青光眼视网膜神经节细胞中表达增强可能在青光眼视神经保护中具有重要作用。     

葡萄糖调节蛋白在急性高眼压大鼠视网膜中的表达及其意义

背景 青光眼导致的视网膜神经节细胞(RGCs)进行性死亡是导致患者视功能损害的主要病理基础,研究表明内质网应激(ERS)参与此过程,葡萄糖调节蛋白78( GRP78)是内质网中的特异性标志物,检测GRP78在高眼压诱导视网膜中的表达对青光眼患者视神经功能保护机制的研究具有重要意义. 目的 检测GRP78在大鼠急性高眼压后不同时期视网膜中的表达情况,探讨ERS在急性青光眼损伤中的作用.方法 56只Wistar大鼠采用随机数字表法分为正常对照组、前房穿刺组、急性高眼压12h组及急性高眼压1、3、7、14 d组,每组8只眼.急性高眼压模型的制作采用前房穿刺灌注生理盐水法,眼压提高至66 mmHg.分别于造模后12h及1、3、7、14 d用过量麻醉法处死大鼠.苏木精-伊红染色法观察各组大鼠视网膜的病理形态学改变;免疫组织化学法检测视网膜中的GRP78蛋白表达的变化;实时定量逆转录聚合酶链反应(RTPCR)法检测视网膜中GRP78 mRNA的表达变化. 结果 正常对照组大鼠视网膜各层细胞排列整齐,急性高眼压后12h视网膜水肿增厚、细胞核肿胀,1d时视网膜形态学异常改变达到高峰,3d后水肿减轻,视网膜变薄.免疫组织化学染色显示正常对照组、前房穿刺组大鼠RGCs层和内核层GRP78蛋白呈弱阳性表达,A值分别为0.195±0.006、0.196±0.005,急性高眼压12h组大鼠GRP78在视网膜的表达明显增强,A值为0.293±0.011,造模后1d时GRP78表达强度达到高峰,A值为0.499±0.039,造模后3d时GRP78表达明显降低,A值为0.268±0.017,与正常对照组大鼠比较差异均有统计学意义(t=0.098、0.304、0.073,P＜0.05),但造模后7d和14 d时GRP78在大鼠视网膜RGCs层和内核层中的表达量接近正常,与正常对照组比较差异均无统计学意义( t=0.002、0.001,P＞0.05).实时定量RT-PCR检测表明,GRP78 mRNA在正常对照组大鼠视网膜的相对表达量( 2-△△CT)为1.011±0.013,眼压升高12 h快速上调,为1.536±0.145,1 d时达高峰,为2.141±0.171,3d时迅速降低,为1.420±0.212,与正常对照组比较差异均有统计学意义(t=0.525、1.130、0.409,P＜0.05),而造模后7d及14 d GRP78 mRNA在大鼠视网膜的表达量明显下降,与正常对照组比较差异均无统计学意义(t=0.020.0.004,P＞0.05).结论 GRP78参与大鼠急性高眼压后视网膜损伤的发生机制,提示通过对ERS过程进行干预可能达到保护急性高眼压眼视神经功能的目的.     

缺氧诱导因子1α在慢性高眼压下大鼠视网膜中的表达

目的了解缺氧诱导因子1((hypoxia-inducible factor-1α,HIF-1α)在实验性慢性高眼压大鼠视网膜中的表达并探讨其在青光眼视网膜损伤中的作用。方法60只大鼠随机分为假手术对照组;高眼压3d组;高眼压7d组;高眼压14d组;高眼压21d组;高眼压28d组。浅层巩膜静脉烧灼法制成慢性高眼压模型,通过逆转录聚合酶链式反应(RT-PCR)方法和蛋白印迹法(Western-blot)研究HIF-1在慢性高眼压下不同时段视网膜中的表达情况。结果HIF-1αmRNA和蛋白在高眼压的视网膜表达较正常视网膜明显提高(P<0.05)。高眼压后7天达到高峰,可维持28天。结论HIF-1参与了慢性高眼压的视网膜的损伤的信号转导过程,为青光眼视神经损伤的缺氧学说提供了直接证据。     

Lack of neuroprotection against experimental glaucoma in c-Jun N-terminal kinase 3 knockout mice

To determine if the absence of c-Jun N-terminal kinase 3 (JNK3) in the mouse retina would reduce retinal ganglion cell (RGC) loss in mice with experimental glaucoma. C57BL/6 mice underwent experimental intraocular pressure (IOP) elevation with a bead/viscoelastic injection into one eye. One-half of the mice were Jnk3 homozygous knockouts (KO) and were compared to wild type (WT) mice. IOP was measured under anesthesia with the TonoLab, axial length was measured post-mortem with calipers after inflation to 15 mmHg, and RGC layer counts were performed on retinal whole mount images stained with DAPI, imaged by confocal microscopy, and counted by masked observers in an image analysis system. Axon counts were performed in optic nerve cross-sections by semi-automated image analysis. Both WT and Jnk3^−/− mice had mean elevations of IOP of more than 50% after bead injection. Both groups underwent the expected axial globe elongation due to chronic IOP elevation. The absence of JNK3 in KO retina was demonstrated by Western blots. RGC layer neuron counts showed modest loss in both WT and Jnk3^−/− animals; local differences by retinal eccentricity were detected, in each case indicating greater loss in KO animals than in WT. The baseline number of RGC layer cells in KO animals was 10% higher than in WT, but the number of optic nerve axons was identical in KO and WT controls. A slightly greater loss of RGC in Jnk3^−/− mice compared to controls was detected in experimental mouse glaucoma by RGC layer counting and there was no protective effect shown in axon counts. Counts of RGC layer cells and optic nerve axons indicate that Jnk3^−/− mice have an increased number of amacrine cells compared to WT controls.     

Neuroprotective effect of latanoprost on rat retinal ganglion cells

Background To investigate the neuroprotective effect of intravitreal administration of latanoprost on retinal ganglion cell (RGC) damage induced by N-methyl-D-aspartic acid (NMDA) or optic nerve axotomy.Methods Using Sprague-Dawley rats, retinal ganglion cell damage was induced by either intravitreal administration of NMDA or optic nerve axotomy. Latanoprost at doses of 0.03, 0.3, 3, 30 and 300 pmol was administered intravitreally before NMDA injection or optic nerve axotomy. Retinal damage was evaluated by counting the number of surviving RGCs retrogradely labeled with fluorogold under the microscope.Results Seven days after the NMDA injury, the number of surviving RGCs was significantly increased at doses of more than 30 pmol atanoprost (846±178 cells/mm² P=0.0166) compared with vehicle control (556±122 cells/mm²). Ten days after the optic nerve axotomy, the number of surviving RGC was significantly increased even at a dose of 0.3 pmol (815±239 cells/mm², P=0.0359) compared with control (462±75 cells/mm²).Conclusions Intravitreal administration of latanoprost has a neuroprotective effect on rat RGC damage induced by either NMDA or optic nerve axotomy, while its pharmacological features are different.     

光相干断层扫描连续监测大鼠慢性高眼压模型视盘神经纤维厚度变化

目的:用光相干断层扫描（OCT）连续观测大鼠慢性高眼压模型视 盘神经纤维层（RNFL）厚度的变化。 方法:选用Wistar大鼠48只，随机分为3组，每组16只鼠32只眼 ，右眼为激光光凝眼，左眼为对照眼。用波长为532 nm氩激光在全麻下光凝右眼小梁网，引 起眼压 慢性、中等程度升高并观测眼压变化。眼压升高后第3、6、9周时用OCT做视盘线性扫描， 计算机自动测量视盘RNFL厚度，然后处死大鼠，将每组8只大鼠右眼做光学切片行组织学 测量RNFL厚度，将另外8只大鼠右眼做全视网膜铺片甲苯胺蓝染色，记数视网膜神经元细胞 密度，将结果进行比较分析。 结果:激光光凝后大鼠眼压缓慢、中等程 度升高，在第3、6、9 周时光凝眼眼压分别比对照眼眼压为显著升高，差异有统计学意义(P＜0.001)。 OCT检查结果显示在3、6、9周时大鼠光凝眼视盘RNFL厚度分别小于对照眼，差 异有统计学意义（P＜0.05）。处死大鼠后组织学测量RNFL厚度，在3、6、9周时，光 凝眼为（64.38±6.54）、（51.47±6.4）、（42.10±6.10）μm，对照眼厚度为（76.23±6.78）、（78.64±6.15）、（77.64±6.63）μm。将两种方法测 得RNF L厚度值进行回归分析，两者变化趋势一致，相关系数(R=0.932，P＜0.001)。全视网 膜铺片甲胺蓝染色结果显示两组视网膜神经元细胞（RGC）密度值差异有统计学意义（P＜0.0 5）。 结论:激光光凝大鼠小梁可以成功建立大鼠慢性高眼压模型；OCT对大鼠慢性高眼压模型视盘RNFL厚度的测量与 在光学显微镜下的测量值变化趋势一致，相关性好；OCT可以连续活体监测大鼠慢性高眼压 模型视盘神经纤维厚度变化，从而了解大鼠青光眼视神经病变的进展。     

Erythropoietin schützt retinale Ganglienzellen und die Sehfunktion nach Ischämie und Sehnervkompression im Rattenmodell

Background

The glycoprotein erythropoietin (EPO) has been shown to be protective in models of neuronal disease and reduced apoptosis of retinal ganglion cells (RGC) after transection of the optic nerve and in glaucoma. In this study we assessed in vivo the properties of EPO on survival of RGC after ischemia and optic nerve compression, as well as on postischemic visual function. Furthermore, the safety of intravitreal injection was assessed.

Methods

In all experiments, EPO was administered intravitreally in male Brown Norway rats. Ocular ischemia was induced by elevating the intraocular pressure for 55 min. The calibrated optic nerve compression was performed for 10 s. RGC were marked stereotactically and quantified by fluorescence microscopy. The retinal function was quantified by electroretinography (ERG) and the whole visual pathway by visual evoked potential (VEP).

Results

EPO (2 and 20 units per eye, n=9–21) increased the survival of RGC after ischemia by 21±21% and 127±31% (mean ± SEM) and after optic nerve compression by 28±12% and 58±13%. With EPO (20 units), postischemic function was increased, in ERG by 71±13% (a-wave) and 75±19% (b-wave) and in VEP by 264±65% (p=0.053). Neither the ERG parameters, nor the VEP, nor the number of RGC differed significantly after intravitreal injection of EPO (5, 50, and 200 units, n=6–7) in healthy eyes.

Conclusion

The combination of toxicological safety and protection of retinal neurons makes EPO a promising drug for ischemic retinal diseases and traumatic optic neuropathy.     

Intrinsic activation of PI3K/Akt signaling pathway and its neuroprotective effect against retinal injury

PURPOSE: The aim of this study was to determine whether the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway can function as a neuroprotective pathway following induced retinal injury. METHODS: The activation of Akt was assessed by immunoblot analysis, and the role of PI3K/Akt pathway was evaluated by TUNEL staining and counting the number of retrogradely-labeled retinal ganglion cells (RGCs) in the whole retina at 168 h after injury with or without PI3K specific inhibitor, LY294002. RESULTS: Akt was induced within one hr and reached a maximum 6 hrs after optic nerve clamping. The activation was observed in the RGC layer including RGCs, the inner plexiform layer, inner nuclear layer, and in the photoreceptor outer segments. The number of surviving RGCs was decreased significantly 168 hrs after injury. LY294002 partially inhibited the activation of Akt, and significantly decreased the number of surviving RGCs as compared with that of injury alone. CONCLUSIONS: These results indicate that the PI3K/Akt signaling pathway is activated intrinsically and has a neuroprotective effect on injured RGCs.     

玻璃体腔注射大麻素HU-211治疗鼠青光眼视神经损伤的研究

目的：评价玻璃体腔内注射大麻素HU-211对大鼠青光眼模型视神经的保护作用,为青光眼视神经损伤治疗提供实验依据。

方法：采用电凝巩膜表面静脉法制作大鼠青光眼模型18眼,随机分为3组：A组分别隔日一次玻璃体腔注射1mg/0.1mL大麻素HU-211,B组隔日一次玻璃体腔注射0.1mL生理盐水,C组为高眼压组。随机选6只对侧眼为空白对照组,每日观察眼压变化情况,用药4wk后处死大鼠,视网膜冰冻切片,HE染色通过视网膜神经元的密度变化评估大鼠慢性高眼压模型视网膜神经元的损伤程度。

结果： B组的凋亡程度及RGC的损伤程度明显高于A组,差异有统计学意义(P<0.05), B组与C组比较差异无统计学意义(P>0.05)。

结论： 玻璃体腔注射大麻素(HU-211)对大鼠青光眼模型视神经视网膜有明显的保护作用。     

Thickness of the lamina cribrosa and peripapillary sclera in Rhesus monkeys with nonglaucomatous or glaucomatous optic neuropathy

Purpose: To measure the thickness of the lamina cribrosa and peripapillary sclera in monkeys with a nonglaucomatous optic nerve damage and to compare that with those of monkeys with glaucomatous optic neuropathy. Methods: The study included 22 monkey eyes (Macaca mulatta) which had undergone a temporary experimental central retinal artery occlusion (CRAO) and seven monkey eyes in which experimental glaucoma was unilaterally produced. We measured histomorphometrically the thickness of the lamina cribrosa and peripapillary sclera. Results: The lamina cribrosa was significantly thicker in the CRAO group than in the glaucoma group (central region: 212 ± 46 μm versus 167 ± 17 μm; p = 0.009). The thickness of the peripapillary sclera at the optic disc border (253 ± 39 μm versus 192 ± 21 μm; p = 0.001) and outside of the optic nerve meninges (408 ± 70 μm versus 314 ± 64 μm; p = 0.006) was significantly greater in the CRAO group. Conclusions: In monkey eyes with a temporary CRAO as a model for nonglaucomatous optic nerve damage, the lamina cribrosa is significantly thicker than in monkey eyes with experimental glaucomatous optic nerve damage. It may suggest that the loss of optic nerve fibres might not be the reason for the thinning of the lamina cribrosa in eyes with advanced glaucoma. The thinner peripapillary sclera in the glaucomatous eyes may suggest that the monkey sclera is more vulnerable to stretching with increased intraocular pressure than the human eye for which no glaucoma‐related lengthening of the eyeball and thinning of the peripapillary sclera have been observed.     

补肾活血中药对大鼠慢性高眼压模型视网膜神经节细胞PI3K/Akt信号转导通路p-Akt表达的影响

目的　观察补肾活血中药对大鼠慢性高眼压（ｅｌｅｖａｔｅｄｉｎｔｒａｏｃｕｌａｒｐｒｅｓｓｕｒｅ,ＥＩＯＰ）模型视网膜神经节细胞（ｒｅｔｉｎａｌｇａｎｇｌｉｏｎｃｅｌｌｓ,ＲＧＣ）ＰＩ３Ｋ／Ａｋｔ信号转导通路ｐ-Ａｋｔ表达的影响。方法　３０只ＳＤ大鼠随机分成３组:对照组、模型组和给药组,模型组和给药组建立ＥＩＯＰ模型。给药组造模后每天予复方丹参片和杞菊地黄丸的混悬液灌胃,每天１次,连续８周,对照组、模型组每天予生理盐水灌胃。记录造模前、造模后即刻、造模后８周的眼压;光镜下观察各组视网膜厚度和ＲＧＣ数量,电镜下观察视网膜超微结构,免疫组织化学法检测视网膜内ｐ-Ａｋｔ的表达。结果　造模后８周眼压与造模后即刻相比,给药组有降低趋势,差异有统计学意义（Ｐ＜０．０１）。造模后８周,给药组与模型组ＲＧＣ数量均低于对照组,差异均有统计学意义（均为Ｐ＜０．０５）;对照组视网膜厚度与给药组相比,差异无统计学意义（Ｐ＞０．０５）,而对照组与模型组相比,差异有统计学意义（Ｐ＜０．０１）;给药组ＲＧＣ数量及视网膜厚度均高于模型组,差异均有统计学意义（均为Ｐ＜０．０１）。电镜下ＲＧＣ超微结构显示,模型组损伤明显,给药组较模型组有所改善。视网膜ｐ-Ａｋｔ表达免疫组织化学结果分析显示,给药组ｐ-Ａｋｔ阳性染色总面积、平均光密度、积分光密度值与模型组相比,差异均有统计学意义（均为Ｐ＜０．０１）,给药组黑度（１３９．９７±１１．７９）虽高于模型组（１３７．９５±６．１３）,但差异无统计学意义（Ｐ＞０．０５）。结论　补肾活血中药用于ＳＤ大鼠慢性ＥＩＯＰ模型,能降低眼压,提高ＲＧＣ数量及改善超微结构,增加视网膜厚度,上调ＰＩ３Ｋ／Ａｋｔ信号转导通路中ｐ-Ａｋｔ的表达。     

眼内窥镜下联合手术治疗晚期新生血管性青光眼的疗效

目的：观察小梁切除术结合眼内窥镜技术下前部玻璃体切割术联合视网膜激光光凝术治疗晚期新生血管性青光眼的疗效。方法：对2012-07/2013-07期间在我院住院的15例15眼伴光感以上视力的新生血管性青光眼患者使用小梁切除术结合眼内窥镜技术下前部玻璃体切割术联合视网膜激光光凝术治疗的临床资料进行回顾性分析。结果：所有患者出院后随访6 mo。15例15眼术后1 wk眼压12.53±3.73mmHg (1mmHg=0.133kPa),较术前眼压58.81±5.91 mmHg 明显降低,差异有统计学意义( P<0.05),术后1mo患者平均眼压18.26±4.31mmHg,术后3mo患者平均眼压17.06±3.65mmHg,术后6mo患者平均眼压16.13±3.66mmHg,患者术后随访各时段的眼压与术前比较,差异具有统计学意义(P<0.05)。术后视力提高2例2眼(13%),无变化者11例11眼(73%),视力下降者2例2眼(13%)。随访中有4例4眼眼压增高,使用1~2种降眼压眼药水及局部按摩眼球使得患者眼压<21 mmHg。术后眼部疼痛症状明显缓解。15例15眼虹膜及房角新生血管均有不同程度回退。术后随访眼部B超检查未见眼球萎缩、脉络膜及视网膜脱离。结论：小梁切除术结合眼内窥镜技术下前部玻璃体切割术联合视网膜激光光凝术治疗晚期新生血管性青光眼,能有效降低眼压,挽救患者残存视力,解除疼痛,为一种较安全而有效的治疗方法。